A chemiluminescence based approach to nucleic acid testing to detect HBV, HCV, and HIV-1 in blood screening

Hepatitis B virus (HBV), hepatitis C virus (HCV), and human immunodeficiency virus (HIV) are life-threatening blood-borne infections that can be unintentionally transmitted in transfused blood products. Here, we optimized a method for detecting HBV, HCV, and HIV-1 in blood screening using magnetic nanoparticles (MNPs) and polymerase chain reaction (PCR)-chemiluminescence. Viral genomes were extracted from donated blood samples using MNPs. The isolated viral DNA and RNA were amplified in a one-step parallel reverse transcription-PCR (RT-PCR). HBV, HCV, and HIV-1 were detected using complementary nucleic acid probes and quantified using a chemiluminescent substrate. 10 422 donated blood samples were tested both with the above method and Roche Cobas TaqScreen MPX Test. The lengths of the amplified PCR products for HBV, HCV, and HIV-1 were 119 bp, 220 bp, and 174 bp, respectively, indicating that the extraction methods successfully separated high-quality viral genomes from the serum samples. The probes and reaction conditions used for the chemiluminescent detection of HBV, HCV, and HIV-1 genomes in unknown samples were empirically optimized. Of the 10 422 blood samples screened, 12 were HBV positive and 2 were HCV positive. These results were consistent with those of a parallel-controlled study using Roche Cobas TaqScreen MPX Test. The assay described here is fast, accurate, and sensitive for detecting HBV, HCV, and HIV, which runs in parallel and has broad implications for blood screening and epidemiological studies.